Synthesis, characterization and potential biomedical applications of N-succinimidyl ester functionalized, polypyrrole-coated polystyrene latex particles

N-Succinimidyl ester functionalized polypyrrole-coated polystyrene latex particles (PS_E-PPyNSE) were prepared by the in situ copolymerization of pyrrole and the active ester-functionalized pyrrole (pyrrole-NSE) in the presence of polystyrene latex particles. Polystyrene microspheres were prepared by emulsion polymerization (PS_E) leading to particles having a diameter of 450 nm. These PS_E particles were precoated with poly(N-vinylpyrrolidone) prior to the in situ copolymerization of pyrrole and pyrrole-NSE. The initial comonomer concentration fractions were 25/75, 50/50 and 75/25 for pyrrole and pyrrole-NSE, respectively. The PPy-coated PS_E particles were characterized in terms of morphology, particle size, electrophoretic mobility and chemical composition. The study of morphology by means of scanning electron microscopy showed roughening of the underlying PS_E particles owing to the addition of PPyNSE, the overlayer thickness of which was estimated to be around 7 nm. Moreover, loading PPyNSE overlayers resulted in a shift of the electrophoretic mobility from –5.31 m cm/V s to a very small but positive value (0.082–0.112 m cm/V s). X-ray photoelectron spectroscopy and IR spectroscopy permitted the detection of pyrrole-NSE repeat units at the surface indicating that pyrrole and pyrrole-NSE did indeed copolymerize. The PS_E-PPyNSE particles were further evaluated as bioadsorbents of human serum albumin used as a test protein. For this study, PS_E-PPyNSE₅₀ particles, synthesized from a comonomer feed ratio of 50/50 in pyrrole/pyrrole-NSE, were used and were shown to attach efficiently human serum albumin macromolecules with a maximum amount of 0.2 mg m^–2.

Formation and morphology of latex monolayers. Computer simulation studies

The results of computer simulations of monolayers created from monodisperse latex particles are presented and discussed. Layers are characterized by the normalized coverage,NC (the actual coverage of the surface related to its maximum possible coverage with particles), and by the average number of neighbors,ANN, calculated as the number of particles being in contact with a given one and averaged over all the particles on the surface. Variable parameters used in simulations include: the rate of particles deposition, the probability of lateral movements, the probability of desorption of particles adsorbed on the surface, the probability of covalent immobilization of adsorbed particles, and the on-sphere slip parameter, OSS (characterizing the scattering of a falling particle on the particles being already attached to the surface). Morphology of monolayers is qualitatively characterized by relations betweenANN andNC. It is shown that for all monolayers formed without adhesion (and without repulsion) between the particles adsorbed on the surface the dependence ofANN vs.NC is described by a characteristic master curve (regardless of the values of probabilities of desorption and lateral movements of particles). For the monolayers created including adhesive forces between the adsorbed particles the plots ofANN vs.NC lie above the master curve, while similar plots obtained for the layers made of particles showing various types of repulsive interactions are always placed below it. Thus, the dependencies ofANN vs.NC, derived from computer simulations, can be used for the determination of the character of the interparticle interactions in the real systems.     

Kinetics of Anionic Polymerization of ε-Caprolactone (εCL). Propagation of Poly-ε=CL-K+ Ion Pairs

Kinetics of the anionic polymerization of ε-caprolactone (εCL) initiated with (CH₃)₃SiO^?K⁺ and carried out in THF solution has been studied in the temperature range from 0 to 20°C by using a calorimetric method. From the kinetic results and from conductometric measurements of the dissociation constant of the living Poly-εCL^?K⁺ ion pairs (K_D ²⁰ ? (4 ± 2) × 10^?10 mol/L), we concluded that at the conditions indicated above and for concentrations of active centers ranging from 10^?3 to 3.7 × 10^?2 mol/L, propagation proceeds on the ion pairs and is disturbed neither by dissociation nor by the formation of higher aggregates. For the polymerization of εCL proceeding on the poly-εCL^?K⁺     

Design of polyglycidol-containing microspheres for biomedical applications

The paper presents a short review on the synthesis, characterisation and selected medical applications of poly(styrene/α-tert-butoxy-ω-vinylbenzyl-polyglycidol) (P(S/PGL)) microspheres. The soap-free emulsion-polymerisation of styrene and α-tert-butoxy-ω-vinylbenzyl-polyglycidol macromonomer (PGL) in water yielded core-shell microspheres with a low particle-diameter dispersity (ratio of the weight average particle diameter and the number average particle diameter). The interfacial fraction of PGL units, estimated by XPS, was in the range of 0–42 mole % depending on the concentration of the macromonomer in the polymerisation feed. The studies of adsorption of model proteins showed that the surface fraction of adsorbed protein was significantly reduced when the PGL interfacial fraction was higher than 40 mole %. The P(S/PGL) particles with covalently immobilised proteins were used for the preparation of photonic crystal assemblies suitable for applications in optical biosensors and the medical diagnostic test for the detection of Helicobacter pylori antibodies in the blood serum.     

Deposition of poly(styrene/alpha-tert-butoxy-omega-vinyl-benzyl-polyglycidol) microspheres on mica plates crossing the liquid-air interface: formation of stripe pattern

Formation of stripelike assemblies of poly(styrene/alpha-tert-butoxy-omega-vinyl-benzyl-polyglycidol) microspheres adsorbed on nonpatterned mica plates moving perpendicularly from suspension of particles through the water-air interface has been observed. It was found that ordered assemblies were formed by capillary forces acting on particles crossing the water-air boundary. At sufficiently high rates of plate movement (i.e., at appropriate dynamic loading conditions) the adsorbed microspheres approaching the water surface begin sliding on the plate, due to capillary forces, in the direction opposite to the plate movement and are kept below the water surface. Plate movement brings new adsorbed particles to the water-air interface, where particles are assembled into aggregates. When particle aggregates are large, the capillary forces cannot overcome shearing forces and the particle assemblies are withdrawn on the plate above the water surface. This process repeated during continuous movement of the plate results in the formation of the quite regularly distributed stripes of adsorbed microspheres. Formation of the regularly distributed particle assemblies depends on concentration of microspheres in suspension.     

Assemblies of poly(styrene/⇌-tert-butoxy-ω-vinyl-benzyl-poryglycidol) Microspheres with different diameters deposited on mica plates

Formation of poly(styrene/α-tert-butoxy-ω-vinyl-benzyl-polyglycidol) microsphere assemblies on mica plates modified with 3-aminopropyltriethoxysilane was investigated. Microsphere assemblies contained two types of particles similar with respect of their chemical structure but with different diameters (D _n = 1000 and 350 nm). Methods of particle deposition included: deposition from water suspension of a mixture of small and large particles on mica plates placed at the bottom of suspension container, deposition of particles from a drop of ethanol suspension (containing large and small microspheres) placed on the mica substrate, deposition of microspheres on modified mica plates crossing the liquid-air interface-sequential deposition of large and small particles, and one-batch deposition from a mixed water suspension of large and small microspheres. Deposition from water suspension containing large and small microspheres on plates placed on the bottom of suspension container yielded assemblies with large particles randomly distributed among the small ones. Fraction of large particles in adsorbed particle assembly was smaller than fraction of large particles in suspension. Particle assemblies prepared by placement of ethanol suspension of large and small microspheres on mica were composed of quite regularly distributed large particles among the small ones. A two step process consisting of withdrawal of mica plate from water suspension of large particles and then on using this plate as substrate in a second step during which the plate was withdrawn from suspension of small particles yielded particle assemblies containing aggregates of large particles randomly distributed among the small ones. Withdrawal of mica plates from water suspension of large and small microspheres resulted in particle assemblies composed of regularly distributed stripes of large and small microspheres. Formation of the described above microsphere assemblies is possible only in case of reversible adsorption of particles.     

Highly hydrophilic surfaces from polyglycidol grafts with dual antifouling and specific protein recognition properties

Homopolymer grafts from α-tert-butoxy-ω-vinylbenzyl-polyglycidol (PGL) were prepared on gold and stainless steel (SS) substrates modified by 4-benzoyl-phenyl (BP) moieties derived from the electroreduction of the parent salt 4-benzoyl benzene diazonium tetrafluoroborate. The grafted BP aryl groups efficiently served to surface-initiate photopolymerization (SIPP) of PGL. In similar conditions, SIPP of hydroxyethyl methacrylate (HEMA) permitted the production of PHEMA grafts as model surfaces. Water contact angles were found to be 66°, 15°, and 0° for SS-BP, SS-PHEMA, and SS-PPGL, respectively. The spontaneous spreading of water drops on SS-PPGL was invariably observed with 1.5 μL water drops. PPGL thus appears as a superhydrophilic polymer. Resistance to nonspecific adsorption of proteins of PPGL and PHEMA grafts on gold was evaluated by surface plasmon resonance (SPR) using antibovine serum albumin (anti-BSA). The results conclusively show that PPGL-grafts exhibit enhanced resistance to anti-BSA adsorption compared to the well-known hydrophilic PHEMA. PPGL grafts were further modified with BSA through the carbonyldiimidazole activation of the OH groups providing immunosensing surfaces. The so-prepared PPGL-grafted BSA hybrids specifically interacted with anti-BSA in PBS as compared to antimyoglobin. It is clear that the superhydrophilic character of PPGL grafts opens new avenues for biomedical applications where surfaces with dual functionality, namely, specific protein grafting together with resistance to biofouling, are required.     

Colloidal liquid crystal type assemblies of spheroidal polystyrene core/polyglycidol‐rich shell particles (P[S/PGL]) formed at the liquid‐silicon‐air interface by a directed dewetting process

A method for the preparation of stripe‐like monolayers of microspheroids is described. The particles were obtained from polystyrene core/polyglycidol‐rich shell microspheres by stretching poly (vinyl alcohol) films that contain embedded particles. The stretching was performed under controlled conditions at temperatures above the T_g of the films and particles. The elongated films were dissolved in water, and the microspheroids were subsequently removed and purified from the poly (vinyl alcohol). The aspect ratio (AR) of the particles, which denotes the ratio of the lengths of the longer to shorter particle axes, was determined by the film elongation. The AR values were in the range of 2.9‐7.7. Spheroidal particles with various ARs were deposited onto silicon wafers from an ethanol (EtOH) suspension. The particle concentration and volume of the suspension were the same in each experiment. Evaporation of the EtOH yielded stripes of spherical particles packed into nematic‐type colloidal crystals and assembled into monolayers. The orientation of the stripes after ethanol evaporation was perpendicular to the triphasic (silicon‐ethanol‐air) interface along the silicon substrate. The adsorbed stripes on the wafers were characterized in terms of their interstripe distance (ID), stripe width, and crystal domain size. Nematic‐type spheroid arrangements in the stripes were the dominant structure, which enabled denser packing of the particles into colloidal crystals than that allowed by the smectic‐type arrangements. Furthermore, the number of spheroids adsorbed per surface unit of the silicon wafers was similar for all ARs, but the width and frequency of the spheroid stripes adsorbed on the wafers were different.     

Properties of poly(styrene/alpha-tert-butoxy-omega-vinylbenzyl-polyglycidol) microspheres suspended in water. Effect of sodium chloride and temperature on particle diameters and electrophoretic mobility

Hydrodynamic and electrophoretic properties of core-shell poly(styrene/alpha- tert-butoxy-omega-vinylbenzyl-polyglycidol) (P(S/PGL)) microspheres suspended in water are described. The microspheres were obtained by surfactant-free emulsion copolymerization of styrene and alpha- tert-butoxy-omega-vinylbenzyl-polyglycidol macromonomer ( M n = 2800, M w/ M n = 1.05). The process yielded microspheres with number average diameter D n = 270 nm and with low diameter dispersity index D w/ D n = 1.01. Shells of P(S/PGL) microspheres were enriched in polyglycidol. Molar fraction of polyglycidol monomeric units in the shells (determined by X-ray photoelectron spectroscopy) was equal to 0.34, which is much higher than the average molar fraction of polyglycidol monomeric units in whole particles of 0.048. Influences of NaCl concentration and temperature on P(S/PGL) microsphere diameters and on their electrophoretic mobility were investigated. It was found that hydrodynamic diameter of P(S/PGL) microspheres, determined by photon correlation spectroscopy, decreased significantly when temperature did exceed a certain value (transition temperature, T t). It has been found that the decrease is more pronounced for higher concentrations of NaCl in the medium. For microspheres suspended in 10 (-1) M NaCl, the hydrodynamic diameter decreased by 8% whereas for the same particles in pure water the diameter decreased by 5.2%. The process of shrinkage was fully reversible. Values of T t for P(S/PGL) microspheres were lower for higher concentrations of NaCl. Adjustment of salt concentration allowed controlling T t in a range from 44.4 to 49.9 degrees C. 13C NMR relaxation time measurements (T 1) for carbon atoms in polyglycidol macromonomer revealed that T 1 did increase with increasing temperature (in temperature range from 25 to 75 degrees C) indicating higher motion of chains at higher temperature. Addition of NaCl did not induce a substantial change of T 1 in the mentioned temperature range. The swelling-deswelling properties of P(S/PGL) microspheres' interfacial layer affected adsorption of P(S/PGL) particles on modified with (3-aminopropyl)triethoxysilane mica. It was shown that the deposition of P(S/PGL) microspheres at 25 degrees C on mica led to formation of two-dimensional crystal-shape assemblies, whereas at 60 degrees C (far above T t = 49.8 degrees C in H2O) the microspheres were randomly adsorbed without formation of colloidal crystal assemblies.     

Mechanism of particle formation and kinetics of the dispersion polymerization of cyclic esters

Pseudoanionic and anionic polymerizations of ε-caprolactone and lactides in 1,4-dioxane:heptane mixtures containing poly(dodecyl acrylate)-g-poly(ε-caprolactone) yield polyesters in form of microspheres. Monitoring partition of active centers between solution and microspheres revealed that particles are formed during initial period, when macromolecules reach their critical masses (ca. 1 000) and became insoluble. Then, propagation proceeds inside of microspheres into which monomer diffuses from solution. Monitoring of variation of the number of particles in a unit volume of reaction mixture with time indicated that after a primary nucleation the delayed nucleation and aggregation are absent. In effect, microspheres with narrow diameter distribution are obtained. Kinetic measurements revealed that in the dispersion pseudoanionic (initiator (CH₃CH₂)₂AlOCH₂CH₃) and anionic (initiator (CH₃)₃SiONa) polymerizations of ε-caprolactone the overall rates of monomer conversion are from 10 to 30 times higher than for the corresponding polymerizations in solution (THF solvent). Analysis of kinetic equations indicated that the observed faster monomer conversions in polymerizations in dispersed systems are due to the high local concentrations of active centers and monomer in growing microspheres.